Summary Discretionary control of renal Na+ transport matches renal Na+ excretion with dietary Na+ intake. Because Na+ excretion can influence blood pressure, disordered renal Na+ transport in many instances causes abnormal blood pressure. Moreover, as we know from the actions of most diuretics and many tubulopathies interdiction of normal renal Na+ transport changes blood pressure. Renal sodium excretion is fine-tuned in response to hormonal signaling in the aldosterone-sensitive distal nephron (ASDN). Within the ASDN, the activity of the epithelial Na+ channel, ENaC, is the principal mediator of Na+ reabsorption. Consequently, modulation of ENaC activity is an important regulator of Na+ excretion and blood pressure. ENaC functions as one final effector of the renin-angiotensin-aldosterone system (RAAS) during the control of blood pressure. Gain and loss of ENaC function, like RAAS, increases and decreases blood pressure by decreasing and increasing renal Na+ excretion, respectively. Emerging evidence supports that there are other physiologically important signaling pathways that function in parallel with the RAAS to fine-tune ENaC activity in the ASDN. Previous R01 funded research from my laboratory demonstrated that a purinergic system intrinsic to the distal nephron regulates ENaC activity through inhibitory paracrine signaling via apical membrane metabotropic P2Y2 receptors in principal cells. Our findings have shown that this purinergic system is quantitatively important to the regulation of ENaC and perhaps consequently, sodium excretion and blood pressure. The latter, though, is only surmised having been tested indirectly and in a cursory manner. Similar to a gain of ENaC function, dysfunction of normal paracrine purinergic inhibition of ENaC is predicted to cause salt-sensitivity and increases in blood pressure as a result of inappropriate Na+ excretion. In contrast, activation of this system is predicted to promote Na+ excretion. The studies proposed in this resubmission test the premise that inhibitory purinergic regulation of ENaC contributes to the fine-tuning of renal Na+ excretion and consequently, regulation of blood pressure. These studies will provide mechanistic understanding and offer a high degree of translation to the human condition by testing the following three aims: 1) Determine if targeted disruption in the ASDN of purinergic signaling increases ENaC activity, decreases Na+ excretion and causes salt-sensitivity; 2) Determine if targeted activation of P2Y2 receptor signaling in the ASDN increases Na+ excretion and can mitigate to some degree forced salt-sensitivity; and 3) Determine if inhibitory purinergic signaling is important for ENaC regulation in the human kidney. It is expected that completion of these studies will elaborate a physiologically important mechanism that contributes to the normal regulation of Na+ excretion; and that when dysfunctional may cause certain forms of salt-sensitivity; and possibly serve as a novel therapeutic target for the treatment of elevated blood pressure.